N = 25) and two (N = 25) and four (N = 16) years later. During the follow-up, the rate constant for brain extraction of glucose (Kglc) declined by 6%–12% mainly in the temporo-parietal lobes and cingulate gyri (p ≤ 0.05), whereas brain acetoacetate extraction (Kacac) and utilization remained unchanged in all brain regions (p ≥ 0.06). Over the 4 years, cognitive results remained within the normal age range but an age-related decline was observed in processing speed. Kglc in the caudate was directly related to performance on several cognitive tests . Peripheral insulin resistance assessed by the homeostasis model assessment of insulin resistance (HOMA-IR) was significantly inversely related to Kglc in the thalamus  and in the caudate , and also inversely related to executive function, attention and processing speed . We confirm in a longitudinal setting that the age-related decline in Kglc is directly associated with declining performance on some tests of cognition but does not significantly affect Kacac.We aimed to longitudinally assess the relationship between changing brain energy metabolism (glucose and acetoacetate) and cognition during healthy aging. Participants aged 71 ± 5 year underwent cognitive evaluation and quantitative positron emission tomography (PET) and magnetic resonance imaging (MRI) scans at baseline ( The assessment of both the brain’s main fuels is particularly informative given that brain ketone uptake remains normal whereas brain glucose hypometabolism is observed in MCI and early AD  or acetoacetate . Our secondary aims were to assess brain structural changes and the relation between change in cognitive performance and changes in brain energy metabolism, brain structure, peripheral energy metabolism and body composition over the same 4-year period.Our overall aim here was to conduct a longitudinal study in a group of cognitively normal older adults, focussing particularly on brain metabolism of both glucose and ketone in relation to cognitive function. Our primary objective was to quantify brain energy metabolism at 2-year intervals over 4 years, with a focus on the brain’s capacity to extract these fuels from the blood, defined as the rate constant for glucose (KN = 8), elevated cholesterol (N = 4), or hypothyroidism (N = 4). At baseline (N = 25), and at the 2-year (N = 25) and 4-year (N = 16) follow-up, participants underwent a general medical examination, complete blood test, body composition analysis, cognitive evaluation and a PET-MRI protocol all within a 6-week period.This study was approved by the ethic committee of the Research Center on Aging—CIUSSS de l’Estrie—CHUS, and was conducted with the informed written consent of all the participants. Of 41 participants in a previous cross-sectional study . After a 60 min wash-out period, an i.v. dose of 191 ± 25 MBq of 18F-FDG was administered and PET images were acquired over 60 min . Repeated arterialize venous blood samples were obtained during the two scans. 18F-FDG PET scans were reviewed by a nuclear radiologist to detect abnormalities.Measurement of brain For all participants and at each time point, T1-weighted (T1w) and FLAIR MRIs were acquired on a 1.5 Tesla scanner . Scan duration was 9.14 min. FLAIR and T1w MRIs were reviewed by a neurologist to detect structural abnormalities as well as potential ischemic stroke. Acquisition parameters were identical at all three time points  as previously described  were quantified according to the graphical analysis method developed by Patlak et al. (−1)]. Subsequently, the CMR is derived from the brain influx K according to the following mathematical equation:Brain k et al. ; based oacac and CMRglc were set to 1.0 and 0.8, respectively.where Cp is the plasma tracer, and LC is the lumped constant. The LC of CMRglc and Kacac obtained with PXMOD tool  were generated using the MIMvista medical program 6.4 .PET images were partial volume-corrected (PVC) using the PVC MR-based solution . The Trail Making Number-Letter sequencing (NLS), Verbal Fluency category-switching and Stroop Color-Word interference inhibition/switching tests from the Delis–Kaplan Executive Function System (D-KEFS) provided information on executive function  and AcAc, were analyzed as previously described  and appendicular lean mass index (arm + leg lean mass/height2), were also calculated  was done by dual-energy X-ray absorptiometry . DXA-derived measures, fat mass index  correction  vs. cognition, brain morphometry vs. cognition, HOMA-IR vs. brain energy metabolism, and HOMA-IR vs. cognition.Results are expressed as mean values with their pooled standard errors, except where specified. The data were analyzed for serial measures using the MIXED procedure of SPSS (IBM SPSS Statistics Version 24.0) with age as the main factor. A linear model between dependent variables and age was used and slope tests for fixed effects were considered significant at an alpha level of 0.05. All neuroimaging analysis underwent a glc over time, mainly in the tempo-parietal lobes and cingulate gyri (p < 0.05 FDR corrected). In contrast, Kacac did not show any statistical difference across the regions during the 4-year follow-up (p ≥ 0.11 FDR-corrected). At the 2 and 4-year time points compared to baseline, there was no difference in regional CMRglc or CMRacac (p > 0.06 FDR-corrected).Participant characteristics are shown in p ≤ 0.01 FDR-corrected).During the 2 and 4-year follow-ups, global brain atrophy (mean ± SD) was 1.2 ± 1.2% and 1.7 ± 1.7%, respectively, or 0.52% annually. Regional analysis of brain volumes showed the same pattern as global brain atrophy . Corticap ≤ 0.05). On the other hand, performance on learning word pairs of the Verbal Paired Associates—Part I (VPAI) improved over time (p = 0.03). After normalization of the results for age , only differences on the Trail Making VS and VPAI tests remained statistically different . Composite z-scores did not show any significant change over the 4 years, except in language which showed improvement .Participants had a mean of 16 ± 4 years education. Overall, cognitive performance at baseline and over the 4-year follow-up remained within the normal range for age , but sigp ≥ 0.19). Participants had an average BMI of 27 ± 0.7 kg/m2 throughout the 4 years. No significant difference was found in body fat parameters throughout the 4 years . However, appendicular lean mass index declined over the 4 years  but remained within the normal range for age . Over the 4-year follow-up, no significant change in plasma metabolites or HOMA-IR index was observed  and the D-KEFS Trail Making Visual scanning tests . Similarly, higher Kacac in the caudate was associated with a higher performance on several components of the D-KEFS Stroop Color-Word, the D-KEFS Verbal Fluency and the WMS-III Verbal Paired Associate (p ≤ 0.04). Higher HOMA-IR was associated with lower Kglc in the thalamus  and caudate . Higher HOMA-IR tended to be associated with lower plasma ketone levels , but no significant relationship was noted between HOMA-IR and regional Kacac (p ≥ 0.51). Lower composite z-scores for executive function , attention and processing speed , were significantly inversely related to HOMA-IR. Better cognitive performance on several tests was positively associated with higher volume of the caudate . No other significant relationship was observed among these different parameters .A significant positive relationship was observed between higher Kglc, the brain’s capacity to extract or acquire glucose, in multiple brain regions. This change in Kglc over time was associated with declining cognitive performance which nevertheless remained within the normal range for age. Several brain regions  demonstrated an annual decline in Kglc averaging 1.5%–3.0% per year. These aging-related changes in brain glucose metabolism were less severe, less extensive and in different regions from the rate of decline in Kglc in the frontal and temporal regions in AD . Hence, we show for the first time in a longitudinal setting that the aging-related decline in brain energy metabolism is specific to glucose and is a physiological change associated with cognitive changes that are considered normal for age.Our main observation is that over the 4 year follow-up, cognitively normal older people have a significant decline in Kglc or Kacac because it is a direct measure of the brain’s capacity to take up these to energy substrates. CMR is the brain’s actual uptake of the tracer which varies with changes in blood glucose or ketones. Plasma glucose does not vary too much in this type of PET study, so CMRglc is usually tightly correlated to Kglc and the two measures show broadly the same differences across the brain or over time. However, because plasma ketones can vary a lot, whereas Kacac does not vary much, Kacac is a better measure of the brain’s actual capacity to obtain ketones when they are available.In the present study, we have focussed on “K,” K18F-FDG PET studies, mostly done by comparing young vs. older healthy adults, have reported that age-related differences in brain energy metabolism mainly affect the frontal and temporo-parietal lobes . Insulin influences three aspects of ketone metabolism: free fatty acid availability from adipose tissue by lipolysis, ketogenesis in the liver and peripheral ketone clearance  compared to year 4 (N = 16). However, our statistical approach based on the linear mixed-effects model is the one specifically recommended for an unequal number of observations (McCulloch et al., This study had several limitations, the main one being the small and unbalanced sample size between baseline and year 2 (both Widespread longitudinal changes in the capacity of the brain to extract glucose occur in healthy, cognitively normal older adults. These changes in brain glucose metabolism exceeded by several fold the annual rate of change in brain volume and were not observed for brain ketone metabolism. Insulin resistance was associated not only with declining cognitive performance but also with lower brain glucose metabolism in this cohort.All datasets generated for this study are included in the manuscript and the supplementary files.SC designed the study. SC, MF and C-AC wrote the protocol. C-AC, EC, SN, ML and ST participated in the PET and/or MR image acquisition and processing. SD and OP performed the longitudinal MRI analysis. C-AC and CH ran the cognitive assessments, and analyzed and interpreted the cognitive data. ID supervised the iDXA acquisition and analysis. MF, VS-P and CV contributed to experimental methodology and biological analyses. CB, NF, TF and ET provided medical supervision and assessments throughout the study. C-AC undertook the statistical analysis. C-AC and SC drafted and revised the manuscript. All co-authors reviewed and commented on the manuscript before submission.SC has received travel honoraria and/or consulted for Accera, Bulletproof, Nisshin Oillio, and Prüvit; he has received test materials for research purposes and/or research funding awarded to the Université de Sherbrooke from Abitec, Ultragenyx and Nestlé. He has a patent application pending on the formulation of a ketogenic drink. SC is President and sole shareholder of Senotec Ltd., company aiming to develop a ketogenic supplement for human consumption. SD is officer and shareholder of True Positive Medical Devices Inc., a company specializing in brain image analysis services. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis and Mycoplasma genitalium. A syphilis test was also performed. Risk factors for STIs were identified by multivariate analysis. The overall prevalence of STIs was 30.4% (34/112) in HIV-uninfected women and 24.4% (10/41) in HIV-infected women. Anogenital Chlamydia trachomatis infection was the most prevalent in both groups of women (20.5% vs 19.5%). There was significant agreement for each STI between self-collected and clinician-collected samples: 91.7%, kappa 0.67, 95% confidence interval (CI) 0.49–0.85 for Chlamydia trachomatis; 99.2%, kappa 0.85, 95% CI 0.57–1.00 for Neisseria gonorrhoeae; 97.7%, kappa 0.39, 95% CI -0.16–0.94 for Trichomonas vaginalis; and 94.7%, kappa 0.51, 95% CI 0.20–0.82 for Mycoplasma genitalium. Women with human papillomavirus had coinfection or multiple infections with other STIs. Risk factors for STIs were being ≤ 25 years old, being employed or a student, reporting a history of STI and having a positive HPV test. A high prevalence of STIs in women in the Tapajós region was found. Cervico-vaginal self-collection is a useful tool for STI screening and can be used in prevention control programs in low-resource settings, such as in northern Brazil.The anogenital prevalence of sexually transmitted infections (STIs) and the use of cervico-vaginal self-collected vs. clinician-collected samples were evaluated for the diagnosis of human immunodeficiency virus (HIV)-infected and HIV-uninfected women in the Tapajós region, Amazon, Brazil. We recruited 153 women for a cross-sectional study (112 HIV-uninfected and 41 HIV-infected) who sought health services. Anal and cervical scrapings and cervico-vaginal self-collection samples were collected. Real-time polymerase chain reaction methods were used for  Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV) or syphilis. This represents approximately one million new infections every day [Mycoplasma genitalium (MG) [The World Health Organization (WHO) estimates that among adults aged 15–49 years there are 357 million new infections each year of either ium (MG) . If untrVery few population-based studies of STI prevalence exist for low- and middle-income countries . In BrazThe Tapajós region is located in northern Brazil and comprises two meso-regions of the state of Pará, namely, the Lower Amazon and the Southwest . The maiThe aim of this study was to determine the anogenital prevalence of STIs and to evaluate the use of cervico-vaginal self-collection versus clinician-collection samples for STI diagnosis in HIV-infected and HIV-uninfected women living in the Tapajós region, Amazon, Brazil.The cross-sectional study was conducted from August 2015 to August 2016 and included nonindigenous, HIV-infected and HIV-uninfected women living in the Tapajós region. A total of 186 women were invited to participate in the study, and 153 of those accepted and were eligible to enter the study. To ensure that the study population was representative, the participants were recruited from nine public health units strategically covering the central, semiperipheral and peripheral points of Santarém. The population of other municipalities search for health care in public health units in the outskirts of town, while the population of Santarém search for health care in public health units downtown. The accessibility and infrastructure of the public health units that had health professionals prepared to perform the sample collections were also considered. We also invited all HIV-infected women at the Santarém Counseling and Testing Center, the only referral center for the care and follow-up of people living with HIV in this region, to participate. The HIV-uninfected women voluntarily sought the public health unit to schedule pap smear collection and were then invited to participate in the study. HIV-infected women who either had an appointment with a healthcare provider without a scheduled gynecological examination or who went to the public health unit to get antiretrovirals were then invited to participate in the survey. The HIV diagnostic testing, HIV viral load determination, and CD4+ T-lymphocyte count were conducted according to the manufacturer’s instructions in the published protocol .The inclusion criteria were being 18 years of age or older , sexually active and able to sign an informed consent form. The exclusion criteria were samples considered unsatisfactory on the pap smear, pregnancy, and use of antimicrobials in the previous 15 days. All women who met all inclusion criteria were considered eligible to participate in the survey.The study was approved by the Universidade do Estado do Pará Ethics Committee Board  and the Instituto Oswaldo Cruz  Ethics and Research Committees. All women agreed to participate in the study and signed an informed consent form in compliance with the Brazilian human ethical guidelines. This study was supported by a previous study in which we evaluated the acceptability of the cervico-vaginal self-collection, and HIV and HPV tests and sociodemographic data are fully described elsewhere . ParticiTrained health professionals of the public health unit collected samples with a brush using anal scraping techniques. Similarly, cervical scrapings were collected with an endocervical brush during the pap smear, and women who agreed to perform the cervico-vaginal self-collection received the collection instructions and an individual collection kit. The self-collection was performed immediately after the pap smear in another office where the woman was alone, and at the end of the procedure, the sample was given to the provider to be labeled/identified and stored . The anaAll collected specimens from scraping and self-collection were immediately stored in ThinPrep Pap Test  at 5°C and transported to the Universidade Federal do Oeste do Pará in Santarém, Pará, for storage and, afterwards, were shipped to the Instituto Oswaldo Cruz, Fundação Oswaldo Cruz in Rio de Janeiro, Brazil. The transportation in Brazil was performed in appropriate polystyrene foam-insulated dry ice boxes with strict temperature monitoring and replenishment of dry ice on every shipment of the blood samples (-20°C to -80°C). All scraping and self-collection DNA samples were transported later to Johns Hopkins University in Baltimore, Maryland, United States of America, in appropriate packaging with strict temperature monitoring (-20°C to -80°C) and replacement of dry ice. All flight transport procedures in Brazil and the USA followed the International Air Transport Association (IATA) international requests for biological materials and were performed by a specialized commercial company (World Courier).DNA extraction from the samples was performed with the QIAamp DNA Mini Kit  and quantified by spectrophotometry using a NanoDrop .A duplex real-time PCR was performed for CT and NG, which was validated at the International Chlamydia Research Laboratory at Johns Hopkins University, utilizing primers and probes from an unpublished assay with sensitivity and specificity of 93.8% and 99.5% for CT and 100% and 100% for NG, respectively, that was validated by comparison with the Roche Amplicor and Hologic Aptima Combo 2 assays. The NG primers were adapted from a previously published protocol . Real-ti2 (25 mM), 2 μl each of the primers and probes (10 μM) and completed with dH2O) and 10 μl of sample DNA to a final volume of 50 μl of PCR reaction mixture. All reactions were performed in a QuantStudio 12K Flex Real-Time PCR System , and the parameters were 95°C for 20 s and 40 cycles of 95°C for 20 s and 60°C for 1 min.All reactions included 40 μl of master mix (25 μl Taqman Fast Advance Master Mix (2X), 2 μl MgClA nested PCR was performed to increase the specificity of HPV DNA detection, with PGMY09 and PGMY11 primers for the first round and GP5+ and GP6+ primers for the second round, as previously described . HPV genTM Syphilis TP test  was used as a point-of-care screening test to detect antibodies to Treponema pallidum (TP) in the serum of HIV-infected women; the test has 100% sensitivity and 100% specificity, according to the manufacturer’s instructions [The Alere Determineructions .P-value ≤ 0.05. For risk factors, the univariate analysis and odds ratio (OR) were calculated, considering the P-value ≤ 0.20. In the multivariable model, only the variables with a P-value ≤ 0.05 remained in the model, and the adjusted odds ratio (aOR) was calculated. Spearman's ρ test was performed to evaluate the linear relationship between the CD4+ T-cell count and STIs. We considered 39 copies/ml for the HIV viral load below the limit of detection (40 copies/ml) for statistical purposes. The positive agreement between cervico-vaginal self-collection and cervical scraping was calculated using the Kappa test with 95% CI and P-value ≤ 0.05. All statistical analyses were performed using Statistical Package for Social Sciences (SPSS) software version 19.0 .Descriptive statistics of the qualitative variables were determined by frequency distribution and the quantitative variables by medians and interquartile ranges (IQR) or means and standard deviation (SD). The chi-square test was used to compare the proportions between the groups for categorical variables and the Mann-Whitney U test for continuous variables, both at 95% CI and A total of 153 women agreed to participate in the study, and overall, they provided 439 samples. Of these, 306 were collected by health professionals using anal and cervical scraping techniques, and 133 were self-collected by participants. The participants were split into two groups: HIV-uninfected women n = 112) and HIV-infected women (n = 41). Overall, 53.1% had three or more pregnancies, 43.8% did not use hormonal contraception, 68.4% had her first sexual intercourse at or before 17 years of age, 54.7% reported four or more sexual partners in her life, 85.2% had pap smears regularly, 58.6% practiced anal sex, only 16.4% used condoms, and 30.5% reported a history of STIs. Additionally, 22.2% had abnormal pap smears, 49.7% had HPV DNA detected in cervical scrapings, and 86.9% performed the cervico-vaginal self-collection . Some re and HIV-P = 0.470). Women diagnosed with cervical lesions and inflammation due to bacterial vaginosis were referred for medical care follow-up.A high prevalence of STIs was identified in the population studied, with occurrences in 30.4% (34/112) of HIV-uninfected and 24.4% (10/41) of HIV-infected women based on positivity in at least one of the three clinical sample types, although no statistically significant difference was observed  and MG (10.7%), while in HIV-infected women, the prevalences for CT (19.5%) and TV (9.8%) were highest, based on positivity in at least one of the three types of clinical specimens . The STIP ≤ 0.05. There was significant agreement for each STI detected in the cervico-vaginal self-collection and cervical scraping groups. The comparisons of the CT DNA, NG DNA, TV DNA and MG DNA evaluations for both collection methods are shown in The cervical scraping samples were used as clinician-collected samples for comparisons with cervico-vaginal self-collection using the Kappa test, with 95% CI, and It is important to note that substantial agreement (kappa> 0.60) was observed for comparisons of CT or NG DNA for all participants, HIV infected women, and HIV uninfected women. MG DNA showed substantial to moderate agreement among HIV-infected and non-HIV-infected women, respectively. The TV DNA presented the worst agreement for the two collection methods .P = 0.879). In The prevalence of each STI and the percentage of coinfections and multiple infections in both groups was 23.2% (26/112) in HIV-uninfected women and 24.4% (10/41) in HIV-infected women . No statFor the HIV-uninfected women, the coinfection rate by decreasing order of prevalence was 50.0% (13/26) for CT-HPV, 26.9% (7/26) for MG-HPV, 7.7% (2/26) for MG-CT-HPV, and 3.8% (1/26) for NG-HPV, for NG-CT-HPV, TV-CT-HPV and for NG-MG-HPV. The group of HIV-infected women had a higher diversity of STIs, and the seroprevalence of syphilis was 9.8% (4/41). Coinfections by decreasing order of prevalence were 40.0% (4/10) for CT-HPV, and 10.0% (1/21) for MG-HPV, for NG-CT-HPV, TV-CT-HPV, TV-HPV, TP-TV-CT-HPV and NG-TP-TV-CT-HPV .P = 0.004).HPV infection was detected in all cases of coinfection and multiple infections, and women with positive HPV DNA tests were compared with those with negative tests. Women with positive HPV tests had coinfection or multiple infections with other STIs, and a statistically significant difference was observed  had only one HPV type and 38.5% (10/26) had multiples HPV types; all HIV-infected women positive for any of the four STIs had multiples HPV types . HPV types and classification by high oncogenic risk identified per woman are detailed below .3, respectively. No statistically significant correlations were found  (P = 0.790).All HIV-infected women were on antiretroviral therapy (ART). The median HIV-1 viral loads among women with negative and positive test results for any of the four STIs were 39  and 39  copies/ml, respectively; the CD4 + T-cell counts were 382  and 416  cells/mm= 0.846) . ConsideP ≤ 0.05): age ≤ 25 years , single status  and alcohol use   and being employed or a student , reporting a history of an STI  and having a positive HPV DNA test  significantly increased the risk of having any of the four STIs . The mul97–5.65) .This is the first study of STI prevalence detected by molecular testing in self-collected samples compared to clinician-collected samples in women living in the Tapajós region, Amazon, Brazil. In the present study, a high STI prevalence was identified, namely, 30.4% in HIV-uninfected women and 24.4% in HIV-infected women, as assessed by positivity in at least one of the three types of clinical specimens. This prevalence is much higher than the 2.1% detected in a previous multicenter study with HIV-infected women  and the In our study, the HIV-uninfected and HIV-infected women were able to easily understand how to perform adequate self-collection, considering the substantial and significant overall agreement of detection for each STI compared to the cervical scraping . This fiAnogenital CT was the most common STI detected, with an overall prevalence of 20.5% for HIV-uninfected women and 19.5% for HIV-infected women. The prevalence of anogenital CT infection was higher than that found in multicenter and isolated studies conducted with HIV-uninfected and HIV-infected women in other low- and middle-income countries , but varAnogenital prevalence of NG identified in the two groups of participants (2.7% vs 4.9%) is in agreement with the range of 0.3% to 0.9% of NG reported for women in Brazil ,31; howeThe seroprevalence of syphilis (9.8%) observed in HIV-infected women was higher than that in high-risk populations for STIs in eastern Africa (8.4%) . The magThere have been very few studies of MG prevalence, and those have assessed specific groups of women . Our finThere was no significant correlation in the univariate or multivariate analysis of HIV infection status and STI positivity and no statistically significant difference between the groups. It is important to note that all HIV-infected women in this study were on cART, and most of them were clinically and immunologically well, with virological suppression. Fortunately, this can be attributed to the efficient public health strategies implemented in Brazil to combat HIV/AIDS, such as the provision of regular medical appointments and treatment for HIV free-of-charge by the Brazilian Ministry of Health ,39. ThisInterestingly, in all cases of coinfections, there was an HPV infection; a statistically significant difference and association in the uni- and multi-variate analyses were observed, suggesting that women with HPV infection are at an increased risk of having another STI. Other studies have shown an association between HPV and other STIs and the development of cervical and anal lesions, with an increased risk for HPV-CT and HPV-MG –42.One limitation of our study was the small sample size in the group of HIV-infected women, which may have been influenced by difficulties with accessibility and the infrastructure of the health units in Santarém, as well as the lack of data in the literature on HIV and STI prevalence in the population studied. To maintain homogeneity between the two groups of women, we decided to match them based on age and the date of diagnosis and survey. We established the criterion of including only HIV-infected women who had been diagnosed with HIV infection during the same study period and in the same age groups as HIV-uninfected women (36.9 ± 13.1). The WHO recognizes that one of the current challenges of STI control strategies is to improve the quality of future estimates and supporting countries to generate their own national estimates. This should be done with more data from studies conducted on the general population with low risk and on key populations. WHO is also exploring alternative methods for generating STI prevalence and incidence estimates, including approaches that use data collected from studies conducted in key populations and the use of new technologies to facilitate diagnosis ,43. The STI surveillance strategies in the Tapajós region should be targeted to the general population of women, independent of HIV-infected status, with a focus on young women and adolescents with a history of STI or HPV infection who are single and use alcohol, especially during their reproductive life. The risk factors found are well established in the literature ,28,40,41Efforts should aim to reduce the burden of STIs for all women, especially at a time when cART reduces the burden of HIV infection, and women can have the ability to talk more about reproductive choices. The mobilization of resources for the implementation of cervico-vaginal self-collection as an alternative for the detection of STIs by rapid diagnostic tests (RDTs) based on nucleic acid amplification assays (NAATs) should be seriously evaluated as a strategy to improve screening programs , consideIn summary, we identified a high prevalence of STIs and a significant positive agreement rate between self-collected and clinician-collected samples in this study. Considering these findings, we suggest the implementation of STI screening and prevention control programs in groups at high risk of acquiring and transmitting STIs, such as young women and HIV-infected women, especially in resource-limited settings and isolated regions of the country. In addition, further epidemiological studies are warranted to investigate the role of HPV with other STIs and the impact of CT and MG on HIV-infected and HIV-uninfected women living in the Tapajós region, Amazon, and other regions of Brazil.S1 Table(DOCX)Click here for additional data file.


Performance improved by 0.11 s (elite) and 0.06 s (sub-elite) in the 20-m flying start and by 0.06 s (elite) and 0.08 s (sub-elite) in the 60-m start block start. Strong post-test correlations were observed between 60-m block start performance and standing five jumps (SFJ) in the elite group and between 20-m flying start and 40-m standing start performance and standing long jump (SLJ) and standing triple jump (STJ) in the sub-elite group. Speed training (ST) shows potential in the reduction of step variability and as an effective short-term intervention program in the improvement of sprint performance.The purpose of this study was to examine the effects of speed training on sprint step kinematics and performance in male sprinters. Two groups of seven elite (best 100-m time: 10.37 ± 0.04 s) and seven sub-elite (best 100-m time: 10.71 ± 0.15 s) sprinters were recruited. Sprint performance was assessed in the 20 m (flying start), 40 m (standing start), and 60 m (starting block start). Step kinematics were extracted from the first nine running steps of the 20-m sprint using the Opto-Jump–Microgate system. Explosive power was quantified by performing the CMJ, standing long jump, standing triple jump, and standing five jumps. Significant post-test improvements ( Past investigations have indicated a significant commonality between sprinting performance (across distances from 20 to 100 m) and explosive power ,4,5. As A mixture of plyometric characters exercises (skips and bounds) and some speed running drills created an alternative sprint-specific running methodology, which represents a high-intensity speed training (ST). ST is treated as a low-volume training strategy and credited with producing significant gains in maximal running velocity . This foRunning velocity is a product of step frequency (SF) and step length (SL) ,14,15 inThe fundamental goal of this study was to investigate the potential effects of acute ST on sprint steps kinematics—how these spatial-temporal characteristics can influence sprint running. A secondary goal was to examine performance including maximal running velocity and power of lower extremity. Sprinters of different competitive levels were recruited to better generalize the results of the study. To mitigate the effects of overtraining, only eight sessions were administered in two 10- and 12-day modules separated with 10 days of recovery. Furthermore, the intervention was administered during the pre-season period to avoid additional confounding variables. A pre-test/post-test design was adopted and included a wide battery of tests and exercises commonly applied in sprint training.Based on this consideration, it was hypothesized that the information of selecting the relevant biomechanical parameters obtained after high-speed training, may indicate the elite of the sprinters (seniors and 100 m < 10.50 s) will show less variability of the sprinting step in the 20-m sprint from a flying start than the sub-elite sprinters (juniors and 100 m > 10.50 s).Seven elite and seven sub-elite healthy male sprinters with a minimum of four to five years of regular sprint training were recruited. Sprinters participating in the experiment were members of the national team. For the purposes of the experiment, they were divided into two groups: Elite and sub-elite. The main division criterion was age  and the personal best times in the 100 m sprint (below 10.50 s—elite). Each participant was medically cleared to participate in sprint training and presented no orthopedic or physiological limitation or injury that could affect sprint performance. All participants had previous experience (minimum six months) in sprint-related strength, speed, and plyometrics training before enrollment. Written consent was obtained after the protocol and procedures were explained in full. Parental consent was also obtained from those individuals under 18 years of age. Sprinters were instructed to maintain their normal intake of food and fluids during the study period. Additionally, they were instructed to avoid any strenuous physical activity 24 h prior to testing as well as refrain from eating 3 h before test commencement. The study design was approved by the Institutional Ethics Committee.The study was performed over a four-week period during the pre-season (April–May) just prior to the outdoor racing season. All participants had been engaged in a standard sprint training regimen (five to seven days of a weekly microcycle) from October to November until inclusion in the study. This training regimen involved general and specialized fitness exercises designed to enhance sprinting technique, increase maximal velocity, and develop strength, power, and endurance. Additionally, many of the participants competed during the indoor competition season (January–February).Eight ST sessions were executed in a 10-day and then 12-day module . The modST focused primarily on developing sprint technique and maximal velocity by various exercises that consisted of various skips, bounds, accelerations, starts, and maximal intensity sprints . TraininMaximal running velocity and lower extremity explosive power were measured one day before the intervention and 48 h after the last training session was completed to ascertain acute training effects. Pre- and post-testing was executed at the same time of the day and under identical conditions. No familiarization was provided as all of the testing protocols were well-known to the participants. A warm-up similar to the one used during the training intervention was administered prior to testing . The test battery included the countermovement jump (CMJ), standing long jump (SLJ), standing triple jump (STJ), and standing five jump (SFJ) to assess explosive power. After 60 min of rest, the participants performed the 20-m sprint (from the flying start) to evaluate sprint step kinematics and then 40 m (from a standing start) and 60 m (using starting blocks) to determine maximal running velocity.The CMJ was used to determine vertical jumping performance. The OptoJump–Microgate optical measurement system  determined contact and flight times with an accuracy of 0.001 s. No restrictions were placed on knee angle during the eccentric phase and the participants were instructed to perform a dynamic double arm swing to attain maximal height. Three trials were separated by 1 min of rest with the highest jump selected for analysis.Horizontal jump performance was measured in the following order: SLJ, STJ, and SFJ. From an erect position with parallel feet placement, the participant executed the SLJ and was required to land on both feet in the long jump pit without falling backwards. Jumping distance was measured to the nearest 1.0 cm. The starting position for the STJ and SFJ was similar. In these jumps, after one or more arm and leg swings, the participant performed the required number of forward jumps with each step on an alternating leg and was required to land on both feet in the half-squat position on a special jumping mattress. Three trials were executed for each jump and the longest distance to the nearest 1.0 cm was recorded. A 2 min rest was provided between each trial and 5–6 min between each jump modality.The reliability of the vertical and horizontal jumping tests was measured using intraclass correlation coefficients (ICC). A posteriori analysis obtained correlations of 0.92 for CMJ jump height and 0.93 for SLJ, 0.93 for STJ, and 0.90 for SFJ jump distances. The large coefficients indicate satisfactory test–retest reliability and may be explained by the extensive familiarization of all participants with executing these jumps.The 20-m flying start, 40-m standing start, and 60-m block start were performed on an indoor track integrated with the Brower Timing TC-System . The photocells were positioned on the track at the start and finish according to the sprint distance . In the x) and standard deviations (SD) were calculated for all dependent variables. Student’s t-test was used to examine pre- and post-test differences in running velocity and jumping performance. Fisher’s least significant difference (LSD) tests were performed post hoc to determine pairwise differences when significant F ratios were obtained. Variability in the nine steps was quantified by calculating the SD and confidence intervals (95%CI). The associations between the performance variables (sprint times and jump distance/heights) were determined by Pearson product–moment correlations. Additionally, hierarchical cluster analysis using Ward’s method was used to determine the linkage distances among the kinematic characteristics grouped as elementary determinants of sprint velocity. A statistical power of 0.90 was determined satisfactory and an alpha level of 0.05 was accepted as statistically significant (denoted in bold font).Means (p < 0.05).Height, body mass, and BMI were similar between the groups. The differences between the elite and sub-elite group for age and personal bests were significant (p < 0.05). Sprint step characteristics are presented in p < 0.05) (p < 0.05). Consequently, the consistent generation of high horizontal velocity in the run-up resulted in greater running velocity with stable SV when sprinting the 20 m distance.Post-test variability in the sprint step characteristics was significant for group and step ( < 0.05) . The intr = −0.76) and SFJ (r = −0.77). In the elite group, a significant relationship was found between the 60-m block start and the 20-m flying start (r = 0.81) and 40 m standing start (r = −0.76). No other significant pre-test correlations were found. Post-test analysis revealed correlations between the 20-m flying start and 40-m standing start and SLJ and STJ performance in the sub-elite group. In the elite sprinters, only the correlation between 60-m block start and SFJ performance was significant (r = −0.87). Many of the horizontal jump tests  were strongly associated with each other, however, no significant pre- and post-test correlations were observed between CMJ with any of the variables in either group.Hierarchical cluster analysis of the grouped variables is illustrated in Significant post-test improvements in 60-m block start times were observed in both the elite and sub-elite sprinters by 1.04% and 1.23%, respectively. Improvements were also noted in 20-m flying start performance by 4.84% and 3.62% in the elite and sub-elite sprinters, respectively. Only marginal improvements  were found for 40-m standing start performance, in which sprinting time improved by 0.98% in the elite group . While ir) from 0.65 to 0.90 between sprinting and drop jump (DJ), SJ, CMJ performance, depending on the sprint distance (20–100 m) and type of jump [r = −0.76) and STJ in the elite (r = −0.85) group. Strong correlations were also observed between 20-m flying start times and STJ performance (r = −0.85) in the sub-elite group but not in the elite group.An important question in this regard is whether ST is more effective than plyometrics training in enhancing explosive power and maximal running velocity. The data from this study can be compared with previous research that used an identical testing protocol to assess the effects of six sessions of plyometrics training . When co of jump ,30,31. TThe secondary purpose of the study was to examine the effects of ST on sprint step kinematics . Analysis of SL in both groups of sprinters was compared with other research where high-performance sprinters were investigated ,32,33,34r = –0.192) between SL and running velocity but a strong correlation between SF and running velocity (r = 0.886). In turn, Hunter et al. (2004) found a strong correlation between sprint velocity and SL (r = 0.73) and only a weak correlation between sprint velocity and SF (r = –0.14). While this contradicts the previous finding, it does confirm Delecluse et al. [Considering both SF and SL, the elite sprinters presented greater SF and slightly longer SL than the sub-elite sprinters . This ree et al.  who usede et al.  examinedPost-test SV increased from 9.10 m/s to 9.26 m/s in the sub-elite and from 9.93 m/s to 9.98 m/s in the elite group. Single sprint step execution in the elite sprinters showed a linear increase across steps one to nine at both pre- and post-test with no changes in SL in step four and seven when compared with earlier steps three and six. In turn, the sub-elite sprinters showed more pronounced variation in SV and SL, which, in turn, may have perturbed the sprinting movement and thus explain the slower running velocity . As SV iNo significant changes were also observed in CT and FT in either group of sprinters . The difTo better understand the effects of ST on the spatial and temporal variability of sprinting, hierarchical cluster analysis was applied. Post-test analysis revealed that the variables in the first cluster show greater Euclidean distances between CT and FT (6.5 at pre-test and 7.5 at post-test). Changes were also observed in the grouping order regarding the CT, which are arranged in the order of the executed steps. Post-test changes in the second cluster (SF and SV) revealed closer sub-cluster linkages (2.5 units at pre-test and 2 at post-test). These results suggest that SF and SV show considerable dependency and may be associated with improvements in running velocity. Similar conclusions can be assumed for CT and FT based on the Euclidean distances.In summary, this study has shown that the application of eight SST sessions are effective in significantly increasing 60-m block start and 20-m flying start sprint performance. Significant improvements were also observed in lower extremity explosive power as ascertained by vertical and horizontal jump testing. Greater increases were observed in the CMJ (mean 7.95% increase) than in the horizontal jumps (mean 2.5–5.3% increase). Increased 20-m running velocity was associated with increases SV, as CT and FT did not change after SST and only relatively linear increases were observed in SL and SF from step two to step eight.Additionally, sprint-speed training can be recommended as an effective short-term intervention to improve sprint performance and lower extremity explosive power, particularly when considering the required training volume of eight sessions (across 22 days). Sprint performance gains can also be optimized by decreasing variability in sprint step kinematics during maximal velocity running in both lower and higher performing sprinters.


Salvadora persica L. root extract (RE) as a bioreductant. The formation of highly crystalline Ag-NPs was established by various analytical and microscopic techniques. The rich phenolic contents of S. persica L. RE (Miswak) not only promoted the reduction and formation of NPs but they also facilitated the stabilization of the Ag-NPs, which was established by Fourier transform infrared spectroscopy (FT-IR) analysis. Furthermore, the influence of the volume of the RE on the size and the dispersion qualities of the NPs was also evaluated. It was revealed that with increasing the volume of RE the size of the NPs was deteriorated, whereas at lower concentrations of RE smaller size and less aggregated NPs were obtained. During this study, the antimicrobial activities of both chemically and green synthesized Ag-NPs, along with the aqueous RE of S. persica L., were evaluated against various microorganisms. It was observed that the green synthesized Ag-NPs exhibit comparable or slightly higher antibacterial activities than the chemically obtained Ag-NPs.Microbicidal potential of silver nanoparticles (Ag-NPs) can be drastically improved by improving their solubility or wettability in the aqueous medium. In the present study, we report the synthesis of both green and chemical synthesis of Ag-NPs, and evaluate the effect of the dispersion qualities of as-prepared Ag-NPs from both methods on their antimicrobial activities. The green synthesis of Ag-NPs is carried out by using an aqueous solution of readily available  The development of various green procedures for the preparation of metallic nanoparticles (NPs) is highly desirable in the current scenario, because of rising environmental apprehensions caused by the climate change ,2. UsualIn this context, the strategies of green chemistry have considerable contributions to the synthesis of metallic NPs . FollowiAmong, various metallic NPs, silver nanoparticles (Ag-NPs) have drawn considerable attention of researchers and scientists because of their wide range of applications in the progress of new technologies in various fields, including, material sciences, nanoscience, electronics, and medicine ,13,14. AMost of the properties possessed by Ag-NPs are largely based on their morphology and size of the NPs. It is extensively reported in various studies that the spherical shaped silver NPs with small size are more effective compared to the other morphologies of NPs ,22,23. RNatural products obtained from different organisms including bacteria, fungi, and plants are proving to be valuable reducing agents for the synthesis of NPs as they exhibit significant chemical diversity yielding wide spectrum of biological activities . These namCD (poly-{6-[3-(2-(3-aminopropylamino)ethylamino) propylamino]}-(6-deoxy)-β-cyclodextrin) system and antibiotics, due to the excellent binding ability of cyclodextrin further resulted in a synergistic improvement of the antibacterial activity of the system. Moreover, the capping of NPs with natural compounds may also facilitate the binding of the synthesized NPs to different bio-surfaces and biomolecules, and may also help NPs cross certain bio membranes [Furthermore, green synthesis may improve the bioactivity of the NPs through the following possible routes. Firstly, it may result in the capping of NPs with biomaterials enhancing their colloidal stability and consequently enhancing their bioavailability ,34. Thusembranes . In factembranes .Salvadora persica L. root extract (RE) as a bioreducing agent for the green synthesis of Ag-NPs. S. persica L. is commonly known as “Miswak” and has rich contents of alkaloids, flavonoids, terpenoids, saponins, and tannins, which makes it attractive for the green synthesis of various materials including graphene [S. persica L. RE towards the green synthesis of metallic NPs, herein we report the preparation of Ag-NPs using this RE both as a reducing and stabilizing agent. Furthermore, during this study, various potential properties of the as-prepared green synthesized Ag-NPs, including their dispersibility and antimicrobial properties were evaluated and compared with that of the chemically synthesized Ag-NPs  prepared by using different volumes of RE was measured as shown in The environmentally friendly preparation of Ag-NPs was carried under reflux conditions using the S. persica L. RE. These samples of SP-Ag were synthesized by using 0.1 mL (SP-Ag-1), 0.5 mL (SP-Ag-2), 1.0 mL (SP-Ag-3), 2.0 mL (SP-Ag-4), and 3.0 mL (SP-Ag-5) of miswak RE taken from the stock solution as mentioned in the experimental section. Notably, the amount of AgNO3 was kept constant in all these experiments. Initial UV analyses of these samples have clearly demonstrated that the formation of Ag-NPs only occurs when a lower volume of RE—between 0.1 and 3 mL  in the S. persica L. RE, which are not present in P. glutinosa PE [These results clearly indicate that inosa PE . When thS. persica L. RE, green synthesized Ag-NPs (SP-Ag-NPs), and chemically synthesized Ag-NPs (C-Ag) as shown in The twin role of the RE as a green reductant and stabilizing agent was established by comparing the Fourier transformed infrared (FT-IR) spectra of pure S. persica L. RE, except slight minimal shifts in a few peaks.This striking resemblance between these two spectra clearly suggests that some of the residual phytomolecules of the S. persica L. RE remained attached on the surface of the green synthesized Ag-NPs. This is further confirmed by comparing the FT-IR spectrum of SP-Ag with that of chemically synthesized Ag-NPs (C-Ag). Since, the chemical synthesis was carried out without using any organic moiety as a stabilizing agent; no functional groups were bound to the surface of C-Ag. Therefore, the FT-IR spectrum of C-Ag does not possess any signal. The FT-IR spectrum of RE exhibit several absorption peaks at different locations including at, 3627 cm−1 (hydrogen bound OH), ~2933 cm−1 , ~2132 cm−1 (C≡C stretch) ~1698 cm−1 (C=O stretch) 1398 cm−1 (C–O stretch) ~1000 cm−1 (C–O) which correspond to various oxygen containing functional groups. Majority of these peaks are also present in the IR spectrum of SP-Ag with some minimal shifts. For instance, all the aforementioned peaks are slightly shifted in the IR spectrum of SP-Ag and appeared at ~3504 cm−1, ~2959 cm−1, ~1663 cm−1, ~1074 cm−1. Therefore, the presence of these peaks in the IR spectrum of SP-Ag clearly points towards the successful dual role of the S. persica L. RE, both as a reducing and capping agent.As shown in S. persica L. RE.Additionally, the SP-Ag was also characterized by X-ray diffraction (XRD) analysis. The diffractogram of SP-Ag exhibited several intense diffractions , which nS. persica L. RE (as a capping ligand) on the surfaces of the NPs.The size and morphology of SP-Ag-NPs prepared with 1 mL of RE (SP-Ag-3) were evaluated by transmission electron microscopy (TEM). S. persica L. RE, before evaporation of solvent (water) from the RE solution, a white powder precipitated out in large amount. The powder is separated out from the solution, which is subsequently evaporated to get the pure RE for the synthesis of NPs. The powder is subjected to various characterization techniques including XRD . The residual organic moieties which were also present on the surface of the Ag-NPs synthesized from P. glutinosa PE [S. persica L. RE. This was further confirmed by comparing the XRD spectra of precipitate obtained from the aqueous solution of RE and the Ag-NPs obtained by using RE. The majority of the additional reflections present in the XRD spectrum of Ag-NPs were also present in the spectrum of precipitated powder from RE aqueous solution. This clearly suggests that the residual amount of precipitated powder not only remained in the pure extract but also bound to the surface of the metallic NPs.The additional reflections in XRD spectrum of Ag-NPs synthesized by inosa PE  did not S. persica L. RE mediated synthesized Ag-NPs are also compared with chemically synthesized Ag-NPs  structure of the Ag-NPs. Hence, the presence of similar reflections in the XRD spectrum of green synthesized Ag-NPs  and Gram-positive (M. luteus and S. aureus) bacterial strains is shown in Ag-NPs synthesized with different volume of Pulicaria glutinosa [S. persica L., a plant with known antimicrobial activity and traditionally used for oral hygiene, was used [S. persica L. exhibited considerable antimicrobial activity at concentrations range of 50–400 mg/mL [Increasing antimicrobial activity of Ag-NPs with the increasing volumes of RE used to synthesize the NPs was also observed in our previous study on lutinosa , mainly was used . However00 mg/mL . In this00 mg/mL ,44.S. persica L. roots, which are widely available in the southern zone of Saudi Arabia, were procured from a local herbal market at Batha, Riyadh, Saudi Arabia. The identity of the plant was confirmed by an expert plant biologist and the details were reserved in our research laboratory with the sample coupon number KSUHZK-302. AgNO3 , tri-sodium citrate and other solvents used in this work were purchased from Sigma Aldrich and were used as purchased.The S. persica L. roots were chopped into small pieces. Subsequently, the chopped pieces (1.3 kg) were drenched in DI water (3 L) and refluxed for ~4–5 h. After reflux the resultant solution was sieved and dried at 50 °C under reduced pressure in a rotary vacuum evaporator. Finally, a dark brownish gluey extract (30.0 g) was obtained, which was stored at 0–4 °C for further use. In order to use this extract for the preparation of Ag-NPs, a stock solution was prepared by dissolving 1.0 g of as-obtained RE in 10 mL of DI water.To begin with, freshly obtained S. persica L. RE (from the stock solution) to 49 mL aqueous solution containing 84.935 mg (0.5 mM) of AgNO3 in a 150 mL flask. The resultant round-bottom flask was equipped with a magnetic stir bar and mounted on a cooling condenser. The mixture was stirred for ~2 h at 90 °C. While stirring, the color of the reaction mixture changed instantly from pale yellow to dim brown and thereafter no color change was witnessed until the end of the reaction (even after two hours). Subsequently, the reaction mixture was allowed to cool down and subjected to centrifugation at 9000 rpm  at room temperature. Thereafter, the acquired product was washed several times (three times) with DI water. Finally, a black powder was achieved, which was dried overnight at 80 °C in an oven. In order to understand the effect of the volume of RE on the preparation of Ag-NPs, several experiments were carried out by taking different amounts of RE. For instance, various experiments were carried out by mixing 0.1, 0.5, 1.0, 2.0, and 3.0 mL of RE (from the stock solution) and 0.5 mmol of AgNO3 (84.935 mg) in particular amount of DI water to make up a total volume of 50 mL. All experiments were executed at 90 °C.The silver nanoparticles were synthesized by the addition of 1 mL of 3 was carried out by using tri-sodium citrate as reducing agent at boiling temperature. To begin with, 50 mL aqueous solution of AgNO3 with 0.001 M was prepared in a 100 mL round bottom flask. The reaction flask was mounted with a cooling condenser and the solution was allowed to reflux at boiling temperature (100 °C). To this boiling solution, 5 mL of 1% trisodium citrate was added slowly. The resultant reaction mixture was kept under constant stirring at 100 °C. Finally, the reaction was stopped when the solution changed to a greenish yellow color. The resultant solution was allowed to cool down and the product was isolated via centrifugation. The Ag-NPs obtained by this method are referred to as C-Ag throughout this article.In order to compare the dispersion quality and the biological activity of both green synthesized and chemically synthesized Ag-NPs. A fresh sample of Ag-NPs was prepared by using tri-sodium citrate as reducing agent. In this method, spherical Ag-NPs were prepared according to the literature . For insOptical measurements were carried out by using UV-visible spectrophotometer . The measurements were carried out in quartz cuvettes by using double distilled water as a reference solvent. In order to perform the kinetic studies, the UV samples were prepared by dispersing 1.0 mL of crude mixture (the mixture collected during the reaction or at the end of the reaction) in 9.0 mL of DI water via sonication for 15 min. Furthermore, UV analysis of pure Ag-NPs was also carried out to confirm their identity. For this purpose, the stock solution of as-prepared Ag-NPs was prepared by dispersing 5.0 mg of pure Ag-NPs in 5.0 mL of distilled water via 1 h sonication. Finally, the sample for the UV analysis of pure Ag-NPs was prepared by diluting 2 mL of stock solution in 8 mL of DI water.To identify the size and morphology of the as-prepared Ag-NPs, TEM analysis was carried out by using JEM 1101 transmission electron microscope . The specimen for the TEM analysis was prepared by dissolving 5 mg of Ag-NPs in 10 mL of ethanol. In order to prepare the grid for TEM measurement, a drop of as-prepared solution is placed on a grid (copper), which was then dried out at 80 °C for 6.0 h in an oven.XRD diffractograms were obtained by using an Ultima IV X-ray powder diffractometer . During these measurements Cu Kα (λ = 1.5418 Å) was used as radiation source.S. persica L. root extract. To measure the FT-IR spectra of pure Ag-NPs, the as-prepared Ag-NPs were frequently washed with DI water, in order to remove any unbound extract or free biomass residue from the surfaces of NPs. The product obtained was centrifuged at 9000 rpm for 30 min and dried out. In order to prepare a pellet for the FT-IR analysis, the purified Ag-NPs were mixed with KBr. Finally, the background correction of the collected IR spectra was performed by using blank KBr pellet as a reference.A Perkin Elmer 1000 FT-IR instrument was used to identify the phytomolecules present in E. coli ATCC 25922, Ps. aeruginosa ATCC 75853, and two Gram-positive bacteria—namely, M. luteus ATCC 10240 and S. aureus ATCC 92213—were grown in sterile Luria broth, nutrient broth, Müller-Hinton broth, and nutrient broth, respectively. E. coli and Ps. aeruginosa were grown on a rotary shaker at 150 rpm at 37 °C while M. luteus and S. aureus were grown at 30 °C. Agar plates were seeded with the mixture of exponentially growing test organisms (105 colony-forming units/mL) with soft agar (0.7% agar). Plates were left standing for 10–15 min at room temperature for the agar to solidify. Wells 8 mm in diameter were punched into the soft agar layer for testing the antimicrobial activity of nanomaterial. Using a micropipette, 100 μg of the Ag-NPs synthesized with different volumes of RE were added to each well on the plates. Water was used as a negative control while 300 μg of the root extract and 100 μg of the chemically synthesized silver nanoparticles were also included in the experiment for meaningful comparisons. Plates were incubated overnight at their respective temperatures for one to two days and zones of inhibition were measured using a ruler. All the experiments were performed in triplicate and the values reported are the mean values of three independent experiments.Antibacterial activity of the engineered nanoparticles was determined by modified Kirby-Bauer disk diffusion method. Briefly, cultures of two Gram-negative bacteria namely S. persica L. RE has been described. With this method, high quality, spherical Ag-NPs were obtained under facile conditions without applying any hazardous chemicals as reductants or stabilizing agents. The NPs were prepared using a very low volume of RE, exhibiting lower size and less aggregation, whereas, the reduction of AgNO3 was stopped at the higher volume of the RE. Furthermore, the study of antimicrobial activities of both chemically and green synthesized Ag-NPs, as well as the pure RE, against various microorganisms has revealed that the S. persica L. RE does not possesses antibacterial activity of its own at lower concentrations. However, it exerts significant effects on the antimicrobial property of the green synthesized Ag-NPs. In addition, the green synthesized Ag-NPs exhibited slightly higher or equivalent antimicrobial activities in comparison to the antimicrobial activities of the chemically synthesized Ag-NPs. Notably, by virtue of its dual nature, S. persica L. RE not only controls the size but also facilitates the stabilization of the Ag-NPs. This significantly enhances the penetration ability of the NPs into the bacterial cell wall and thus results in the improved microbicidal activity of RE capped Ag-NPs. Therefore, due to the high oxidizing ability, low cost, and abundancy, S. persica L. RE has great potential for the reduction of various other metal salts and can be possibly applied for the synthesis of various metallic NPs and other functional nanomaterials.In this study, an environmentally friendly method to the preparation of Ag-NPs using


However, the DDW- and DAC-based collagen scaffolds displayed significant differences in their physical properties  and swelling behaviors. In particular, the weight losses induced by mechanical stimulation reflected the high degradation of DAC-collagen scaffolds. In cell culture experiments using adipose-derived stem cells (ADSCs), the characteristics of mesenchymal stem cell (MSC) did not change in both DDW- and DAC-collagen scaffolds for 10 days, although cells proliferated less in the DAC-collagen scaffolds. Our results suggest that the elimination of acetic acid residue from isolated collagen is recommended to produce collagen scaffolds that provide a stable environment for cells and cell therapy-related applications.Various methods for isolation of type I collagen using acids, bases, enzymes, and their combinations have been applied. However, a lack of standardization exists among type I collagens isolated by various approaches. Consequently, in this study, we assessed the influence of acetic acid residue on type I collagen isolated by pepsin-acetic acid treatment, the fabrication of collagen-based porous scaffolds, and the seeded cells on collagen scaffolds. Unlike the isolated collagen dialyzed by deionized water (DDW), collagen dialyzed by 0.5 M acetic acid (DAC) exhibited structural and thermal denaturation. Both DDW- and DAC-based porous scaffolds at all collagen concentrations (0.5, 1 and 2%  Type I collagen is considered one of the most useful biomaterials and is used extensively in the field of tissue engineering ,2,3,4. AHowever, a lack of standardization exists among type I collagens isolated by various approaches . SpecifiIn this study, we aimed to determine the influence of acetic acid residue on the characteristics of isolated collagen. Moreover, to elucidate the influence of acetic acid residue on collagen-based porous scaffolds and the culturing cells on the scaffolds, we intended to demonstrate the relationships between the material used for scaffolds and the seeded cells.For this purpose, we isolated type I collagen from porcine skin by pepsin-acetic acid treatment and dialyzed it against deionized water (DDW) and 0.5 M acetic acid (DAC). The structural and thermal properties of both DDW- and DAC-collagens were confirmed by comparison to those of commercially available type I collagen. Three different concentrations of isolated collagen were used to fabricate collagen-based porous scaffolds, and their morphological and physical characteristics as well as swelling and degradation behaviors were analyzed. Adipose-derived stem cells (ADSCs) were seeded on the fabricated scaffolds, and the relationships between the collagen scaffolds and the seeded cells were evaluated by analyzing the genetic maintenance and proliferation of the seeded cells.Porcine skin, used in food, was purchased from an open market . It was chosen as the raw material for type I collagen isolation due to its high similarity to human skin ,15. Glacg for 10 min), the supernatant was salted out by 640 mL of 5 M NaCl solution. The precipitate was suspended in 100 mL of ethanol with stirring for 24 h, after which it was recovered by centrifugation and then re-suspended in 50 mL of ethanol. After centrifugation, the precipitate was suspended in 50 mL of 0.5 M acetic acid and then transferred to a dialysis tubing cellulose membrane  and dialyzed against 900 mL of 0.5 M acetic acid for 3 days by changing the solution every 12 h. The molar concentration of acetic acid in the surrounding solution was measured by titration with 0.1 M sodium hydroxide solution. When the molar concentration of acetic acid in the surrounding solution was constantly sustained at 0.5 M for 72 h, the final collagen was referred to as DAC. Collagen that was dialyzed against deionized water (DW) was referred to as DDW when the molar concentration of acetic acid in the surrounding solution was less than 0.0001 M at 72 h. The resulting collagen solutions were frozen at −20 °C and then lyophilized.Type I collagen was isolated by acetic acid-pepsin treatment, as described previously . The isow/v Coomassie brilliant blue R-250, containing 50% v/v methanol and 10% v/v acetic acid, and destained with 40% v/v methanol and 10% v/v acetic acid. A high molecular weight protein marker  was used to estimate the molecular weight of the proteins.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed using the discontinuous Tris-HCl/glycine buffer system, according to the method of Laemmli . The pro−1.Raman spectroscopy  was carried out using a 532-nm laser at a power of 50 mW with an exposure time of 10 s. The Raman spectra were analyzed after normalization of peak intensities to the amide I band at around 1660 cmw/v collagen solution was poured over a cover glass (18 × 18 mm2) and placed in a refrigerator at 4 °C to dry. The high-resolution XRD system  was operated at 45 kV and 200 mA using CuKα radiation (λ = 0.15418 nm).The spacing of the collagen fibers and phase identification of the two isolated collagens (DDW- and DAC-collagen) and the commercial collagen were determined using thin-film XRD. To fabricate the thin films, 0.5 mL of 1% 2 gas.The thermodynamic properties of DDW-, DAC-, and the commercial collagen pellets  in the dehydrated state were evaluated by thermal gravimetric analysis (TGA) and differential scanning calorimetry (DSC), performed using a simultaneous thermal analyzer . The sample was heated from 20 to 500 °C at 1 °C/min in Nw/v) were prepared from DDW and DAC and then homogenized in 50-mL conical tubes by vortexing for 5 min at 4 °C. Air bubbles were removed from the solutions by centrifugation at 3000× g for 3 min. Finally, 380 μL of each collagen solution was placed into one well of a 24-well plate, frozen at −20 °C, and lyophilized for 1 day each. For cross-linking, the fabricated porous scaffolds were immersed in 50 mM EDC solution in 95% v/v ethanol at room temperature for 24 h . Th. Th29].  > 0.05) .water/gdry scaffold for DDW- and DAC-collagen scaffolds, respectively, p < 0.001) at 0.5% w/v collagen and a 1.03-fold higher swelling ratio at 1% w/v collagen . In contrast, at 2% w/v collagen, the DDW-collagen scaffolds exhibited a 1.07-fold lower swelling ratio than the DAC-collagen scaffolds  (The DDW-collagen scaffolds exhibited a 1.53-fold higher swelling ratio than the DAC-collagen scaffolds (107.15 ± 1.01 and 70.07 ± 1.29 g < 0.01) .w/v) and days 1–4 (0.5% w/v), with no marked weight loss after 4 days. However, at 0.5 and 1% w/v, the DAC-collagen scaffolds showed noticeable weight loss during days 0–1 and 7–14. The differences in weight loss between DDW- and DAC-collagen decreased as the collagen concentration increased at day 14 .As seen in w/v collagen and the lowest proliferation at 2% w/v, while only the 0.5% w/v DAC-collagen scaffolds exhibited decreasing proliferation during days 7–14  were the most intense component of the amide I band. Yet, the unordered structures represented by the peak at 1647 cm−1 and the low 1240 cm−1/1445 cm−1 ratio suggested the denaturation of DAC-collagen [−1) for DAC- than for DDW-collagen (3312 cm−1), indicating the weakening of hydrogen bonds in the triple-helical structure of DAC-collagen [While acetic acid  can be used exclusively to isolate collagen, only partial hydration of the collagen is achieved, and intermolecular cross-linked collagen fibrils are not cleaved ,14. In ccollagen ,24. In acollagen . Likewiscollagen .Similar to the structural stability results, the thermal stability of DAC-collagen was lower than that of DDW-collagen; acetic acid residue attributed to an additional weight loss for DAC-collagen (in the 1st weight loss region) and resulted in DAC-collagen exhibiting the lowest proportion of pure collagen (in the 2nd weight loss region). In the DSC thermograms, a weak denaturation enthalpy in DAC-collagen indicated that acid residue induced strong thermal disruption of collagen and leaded to the formation of random chain of gelatin .Porous scaffolds that resist enzymatic degradation can be fabricated using three broad steps, namely, freezing, lyophilizing, and cross-linking ,36. The EDC optimally reacts with carboxyl groups under slightly acidic conditions (pH ~ 5.5) ,41,42. Iw/v scaffolds, a highly flexible structure due to a low collagen concentration is believed to facilitate the expansion of the scaffold network structure. Consequently, the scaffold can hold a large amount of water (DDW-collagen scaffold). However, despite a high flexibility, because of low structural stability, the scaffold cannot retain the absorbed water (DAC-collagen scaffold). Conversely, at a high collagen concentration (2% w/v), the porous network structure showed low expansion, and therefore its water absorbance ability decreased. Particularly, the lowest swelling behavior, which was evident in the 2% w/v DDW-collagen scaffolds, indicated they had a relatively rigid structure and, accordingly, a high structural stability.At all concentrations, the DDW-collagen scaffolds were heavier and thicker than the DAC-collagen scaffolds, and the volume of the DDW scaffolds (about 200–350 μL) was either larger than or similar to that of the DAC scaffolds. These physical characteristics of DDW-collagen scaffolds are associated with their high structural stability. As observed from the swelling degree of the 0.5% w/v DAC-collagen scaffolds was 6.22-fold higher than that of 0.5% w/v DDW-collagen scaffolds, indicating that the scaffolds with a highly flexible structure but low structural stability had low resistance to mechanical stimulation.In the degradation behavior of scaffolds, there were no significant differences among the scaffolds of the DDW group (at day 14), In contrast, weight loss increased with decreasing collagen concentrations in the DAC group (at day 14). In particular, the weight loss of 0.5% w/v collagen concentration and the lowest proliferation at 2% w/v. The combined effect of high volume, high swelling capacity, and low degradation behavior (1% w/v scaffolds) may induce proliferative potential, whereas markedly reduced swelling capacity (2% w/v scaffolds) may reduce this potential. The 0.5% w/v DAC-collagen scaffolds displayed a decrease in cell proliferation from day 7 that may be linked to the weight loss that occurred when degradation restarted.The high porosity and pore interconnectivity of both DDW- and DAC-collagen scaffolds reflected their amenity to cell infiltration and cell proliferation ,45,46. AThe RT-PCR data suggest that the ADSCs maintained the characteristics of MSCs in all scaffolds for 10 d; there was no marked influence of dialysis medium or collagen concentration on the expression of MSC or pluripotency markers, and although the EMT makers were variably expressed, these results did not suggest transdifferentiation of the ADSCs. However, the low volume, thickness, and swelling capacity and high degradation behavior of the DAC-collagen scaffolds may be critical for stable cell behaviors. Thus, DDW-based collagen scaffolds may be more suitable for supporting the long-term culture of cells.In this study, we found that the structural and thermal characteristics of the isolated collagens were dependent on the dialysis medium and that acetic acid induced collagen denaturation. At the same concentration, DDW- and DAC-collagen scaffolds had comparable pore morphologies and were highly porous. However, the acetic acid associated with DAC promoted scaffolds with larger pore sizes and lower free amine indices compared to those of DDW-collagen scaffolds. The physical characteristics of the scaffolds indicated a higher structural stability for DDW-collagen than for DAC-collagen scaffolds. DDW-collagen scaffolds exhibited the ability to absorb and retain large amounts of water and had high resistance to weight loss by mechanical stimulation. Both DDW- and DAC-collagen scaffolds showed the potential to maintain MSC characteristics of ADSCs, but the proliferative activity of ADSCs was higher in the DDW-collagen scaffolds. These results suggest that the elimination of acetic acid residue from isolated collagen is recommended to produce collagen scaffolds that provide a stable environment for cells and cell therapy-related applications.


Leukocyte and neutrophil counts were also higher for LVR patients . For metabolomic analysis, sphingomyelin C20:2 and symmetrical dimethylarginine were higher for LVR patients, but did not reach significance after the correction for the alpha risk. The metabolomic approach did not discriminate patients with and without LVR. However, common parameters that focus on infarction severity, such as infarct size and inflammatory markers, differed between the groups.Left ventricular remodeling (LVR) occurring after ST-segment elevation myocardial infarction (STEMI) is frequent and severe. We present a metabolomic approach as an attempt to reveal unknown biomarkers associated with post-STEMI LVR. Out of 192 consecutive patients with successfully revascularized STEMI, 32 presented LVR and were clinically matched with 32 no-LVR patients. They underwent cardiac magnetic resonance at baseline, three months and 12 months. Blood samples were collected during index hospitalization. Creatine kinase (CK) peak and inflammatory markers were higher for LVR patients compared to no-LVR patients (mean 3466 ± 2211 and 2394 ± 1615 UI/L respectively,  In recent decades, despite improvements in the management of myocardial infarction, left ventricular remodeling (LVR) has remained a severe and common issue, estimated to occur in up to 30% of cases following ST-segment elevation myocardial infarction (STEMI). LVR can lead to chronic heart failure, which is one of the primary causes of death worldwide . AlthougMetabolomics can be defined as the detection, quantification and identification of low molecular weight molecules of the metabolism . In compp = 0.01).Overall, patients were 58 ± 9 years old, 81% were male and 53% had anterior infarction. The mean left ventricular ejection fraction (LVEF) was 47.8 ± 9% and the mean infarct size was 16.1 ± 11.1% of the total left ventricle (LV). LV volumes were assessed by cardiac magnetic resonance (CMR). LVR occurred in 32 patients (16.7%) from a cohort of 192 STEMI patients. They were matched with 32 no-LVR patients for the purpose of the study. LVR patients presented more history of hypertension than no-LVR patients  (p = 0.020) (p = 0.028) and neutrophil count  (Creatine kinase (CK) peak was significantly higher among LVR patients than no-LVR patients  A. Simila= 0.020) B; leukoc< 0.001) C. By allHowever, there was no association between LVR and N-terminal prohormone of brain natriuretic peptide (NT-proBNP) or creatinine .After validation of the kit plate based on QC samples, 142 (75.5%) metabolites were kept for the statistical analysis: Free carnitine and 11 acylcarnitines, 21 amino acids, 12 biogenic amines, 12 lysophosphatidylcholines, 70 phosphatidylcholines, 14 sphingomyelins and the sum of hexoses.p = 0.013 for sphingomyelin C20:2 and mean 0.57 ± 0.15 µmol/L vs. 0.52 ± 0.07 µmol/L respectively, p = 0.037 for SDMA), but did not reach significance after the correction of the alpha risk. Likewise, without reaching significance after Benjamini correction, lysophosphatidylcholine C17:0, phosphatidylcholine C40:6 and C42:3 were lower for the LVR group than for the no-LVR group . Paired principal component analysis (PCA) showed no outliers or grouping of patients according to their metabolic profile  were higher for the LVR group than for the no-LVR group , which was not confirmed after the risk alpha correction. There is some interest in continuing to assess the role of SDMA in larger cohorts.Finally, endothelial metabolism could be affected. In fact, Lorin et al. showed that SDMA—an enzyme involved in oxidative stress by inhibiting the activity of the endothelial nitric oxide synthase—was a marker of left ventricular dysfunction and in-hospital mortality after a myocardial infarction . In a coOur study has several limitations: (1) The small sample size presents a limitation that we tried to constrain partially through the one-to-one matching; (2) Among the 40,000 metabolites currently known, our targeted analysis focused on only 188; and the analysis was performed at a single timepoint (48 h after STEMI onset); (3) The selected threshold for defining LVR, as an increase of 10% of the end-systolic volume, may seem low but is below interobserver variability in MRI  and our From January 2006 to December 2015, 192 patients presenting with their first STEMI, successfully managed within the first 12 h of the onset of chest pain by primary angioplasty with stent implantation, were included in a prospective cohort . The non-inclusion criteria were: Age < 18 years; history of myocardial infarction or coronary bypass grafting; cardiogenic shock on admission; severe comorbidities limiting life expectancy, and contraindications to CMR . CMR was performed at admission (day 6 ), at the 3-month  and the 12-month follow-up . This study was conducted in accordance with the principles of the Declaration of Helsinki and was approved by the hospital’s ethics committee . All patients gave written informed consent.Clinical data including medical history, baseline characteristics and clinical presentation on admission were prospectively collected during the index hospitalization and at the 3 and 12-month follow-ups.32 patients who presented LVR were matched with 32 no-LVR patients with regard to age, sex, left ventricular ejection fraction, infarct size, culprit artery and time to reperfusion.®, Laboratoires Guerbet, Roissy-Charles de Gaulle, France) and after a recuperation-inversion sequence. Late gadolinium enhancement images were acquired for short-axis and long-axis four-chamber views, 12 min after the injection. In an isolated sequence, optimal inversion time for annulling the normal myocardial signal was determined. The parameters applied for the sequences were: Repetition time = 4.9 ms, excitation time = 1.9 ms, flip angle = 15°, slice thickness = 8 mm, and spatial resolution = 1.35 × 1.35 × 8 mm3.CMR was performed using a 3 Tesla image scan . Images were transferred to a workstation for analysis and calculation . Steady-state free precession sequences were used to produce cine-CMR images for multiple short-axis and long-axis two, three and four-chamber views. To cover the entirety of the left ventricle, contiguous 8mm-thick slices were required. First-pass perfusion images were obtained during the injection of gadolinium contrast agents at a dose of 0.2 mmol/kg , left ventricular end-diastolic volume (LVEDV), left ventricular ejection fraction and infarct size were calculated using cine-CMR images. Endocardial and epicardial borders were outlined manually on end-diastolic and end-systolic frames, excluding papillary muscles and the trabeculae.Infarct size was quantified on late gadolinium enhancement imaging using the “full-width at half maximum” method.LVR was defined as an increase of at least 10% of LVESV during the follow-up .Blood samples were obtained during the index hospitalization  and were stored at −80 °C.® Absolute IDQ p180 kit . This kit, combined with a QTRAP 5500 mass spectrometer , enabled quantification of up to 188 different metabolites of 6 different biochemical families. Their essential functions are structural and metabolic, and some of them are involved in signaling pathways. The full list of individual metabolites is available at http://www.biocrates.com/products/research-products/absoluteidq-p180-kit.Targeted quantitative metabolomic analyses were carried out using the BiocratesAll reagents used in this analysis were of liquid chromatography mass spectrometry (LC-MS) grade and purchased from VWR  and Merck . Sample preparation was performed following the Kit User Manual. Briefly, 10 microlitres of serum was extracted in a methanol solution using ammonium acetate after drying under nitrogen flow and derivatizing with phenyl isothiocyanate for the quantitation of amino acids and biogenic amines. The extracts were finally diluted with MS running solvent before flow injection analysis (FIA) and LC-MS/MS analysis. One blank sample , three water-based zero samples , and three quality control samples were also added to the plate of the kit. A seven-point serial dilution of calibrators was added to the plate of the kit to generate calibration curves for the quantification of amino acids and biogenic amines.Before statistical analysis, the raw data were examined in order to eliminate metabolites not measured accurately enough, i.e., metabolites with a concentration below the lower limit of quantitation (LLOQ) or above the upper limit of quantitation (ULOQ). When more than 20% of the concentration values were not measured accurately enough, the metabolite was not considered for further statistical analysis.Routine biomarkers, analysed using routine automats  were: NT-proBNP ; CRP ; High-sensitive cardiac troponin I (Abbott); creatine kinase (Abbott) and creatinine (Abbott).t-test for matched data. Then, Benjamini correction was applied. A p-value ≤0.05 was considered statistically significant.The quality of our paired dataset granted us to perform paired analysis. Unpaired analyses were run afterwards to corroborate the results. All statistical tests were generated by R Software, Version 3.1.1 . The data are expressed as mean ± standard deviation or median with IQR for continuous variables and as frequency, with a percentage for categorical variables. We used the Shapiro-Wilk test to determine the parametric nature of the variables. The Mann-Whitney test was used for non-parametric variables and the Student’s test for parametric variables. For matching patients, comparisons of non-parametric quantitative variables were performed using the Wilcoxon signed-rank test and parametric variables with the Student’s p-value for the test set were calculated. This p-value measures the probability, for a given AUROC, that the corresponding model was the mean model, randomly predicting patient status (remodeled or not) with no added information based on the metabolic profile. Since AUROCs and p-values cannot be considered as being normally distributed, median values instead of means were used to decide whether PLS models separate these two groups satisfactorily. A cut-off of 0.8 and 0.05 was selected for the median AUROC and median p-value, respectively. Global performance of pPLS on our data was considered as satisfactory only if median AUROC ≥ 0.8 and median p-value ≤ 0.05. In this case, variable selection was conducted based on the variable importance for the projection (VIP) and loading parameters. VIP values summarize the importance of each variable for the PLS-DA model, whereas the loading values are indicators of the relationship between the y vector containing the class information  and variables in the X matrix . Variables with a VIP value greater than unity are important for group discrimination. Multivariate analysis was performed using the R package mixOmics (http://www.Rproject.org).An unsupervised analysis was conducted using paired PCA to detect similar groups of patients and samples displaying an atypical metabolite profile (outliers). Paired PLS-DA was performed to discriminate between LVR and no-LVR patients based on their metabolic profile. PLS-based models are prone to overfitting, sometimes leading to overly optimistic models with poor predictive capabilities. To test the predictive capabilities of PLS-based models, data are often divided into training (~2/3 of samples) and test (~1/3 of samples) sets. Models are then constructed using the training set and their predictive capabilities are tested using the test set. A number of parameters are used to measure the performance of the models in the test set; however, the area under the Receiver Operating Characteristic is one of the most common parameters used because it is parsimonious, easy to interpret and enables performance comparison with other classification methods unrelated to PLS. Training set-based models with AUROC ≥ 0.8 on the test set are considered to be good predictive models whilst AUROC ≥ 0.9 is indicative of very good predictive models with a low degree of overfitting. Training and test set partition is usually done only once and randomly, based on the hypothesis that the probability of obtaining training and test sets representative of the entire population is much higher than that of obtaining training or test sets with extreme configurations that are totally different from the initial population. However, when dealing with small sample sizes, the probability of obtaining less representative training or test sets is far from negligible and could lead to misleading conclusions based on the training/test set partition strategy. Considering this issue and by allocating 22 samples (~2/3) to the training sets and 10 samples (~1/3) to the test sets in each group (LVR and no LVR), a total of 64,512,240 models are possible (The usual parameters that focused on infarction severity, such as infarct size and inflammatory biomarkers, differed between patients with and without LVR. The metabolomic approach performed soon after the onset of STEMI failed to discriminate between these patients.


Prostate cancer is a common malignancy of the male genitourinary system that occurs worldwide. The current research aims to investigate caveolin-1 expression in prostate cancer tissue and its relationship with pathological grade, clinical pathologic staging, and preoperative prostate-specific antigen (PSA) levels.From January 2012 to December 2014, samples from 47 patients with prostate cancer who had received transurethral prostatic resection (TURP) and 20 patients with benign prostatic hyperplasia were collected at the First Affiliated Hospital of Guangxi Medical University. Caveolin-1 was detected by streptavidin-perosidase (SP) immunohistochemical staining in pathological tissue slices. The results were statistically analyzed for pathological grade, clinical stage, and preoperative PSA level.P < 0.05), and caveolin-1 expression was significantly different among the pathological grades of poorly, moderately and well-differentiated prostate cancer (P < 0.05). The difference in caveolin-1 expression was significant for different clinical stages (T1-T2 and T3-T4) of prostate cancer (P < 0.05). The difference in caveolin-1 expression was not significant among samples with different preoperative PSA levels  (P > 0.05).The expression of caveolin-1 was significantly higher in prostate cancer samples than in benign prostatic hyperplasia samples (Caveolin-1 is closely related to the pathological grade and clinical stage of prostate cancer after transurethral surgery, and it may be a novel tumor marker for prostate cancer. The expression of caveolin-1 is not associated with preoperative serum PSA levels. Prostate cancer is a common malignancy of the male genitourinary system. In recent years, there has been a continuous rise in the incidence of prostate cancer cases, and it is becoming the first cancer to threaten male health , 2. ScreAt present, the pathogenesis and relevant molecular biological basis of prostate cancer are not yet fully understood . Recent Caveolin (Cav) is a special vesicle-like structure on the surface of the cell membrane and is an important structural-functional protein. It has been found that mammals have three caveolin genes . Caveolin-1 and caveolin-2 are widely present in a variety of normal human cells, and caveolin-3 is expressed specifically in muscle. Caveolin interacts with specific lipids, such as glycosphingolipids or cholesterols, thereby leading to the formation of caveolae. In the sub-cell, caveolin-1 is positioned at the cytoplasm, mitochondria, caveolae, and other related secretion channels . CaveoliA large number of studies have shown that caveolin-1 is expressed at a low level in most tumors and is present as a tumor suppressor gene . HoweverIn this study, caveolin-1 was detected in pathological tissue slices of prostate cancer by streptavidin-peroxidase (SP) immunohistochemical staining. Statistical analysis was performed on the immunohistochemistry results for the tissue slices to investigate the correlation of caveolin-1 with the prostate cancer pathological grade, clinical stage, and preoperative serum PSA level to provide new information for the clinical diagnosis and treatment of prostate cancer.Inpatients who received transurethral resection of prostate in the Urology Department of the First Affiliated Hospital of Guangxi Medical University between January 2012 and December 2014 were included in this study. The inclusion criteria were patients who had undergone surgical resection for prostate cancer and had benign prostatic hyperplasia that had been diagnosed pathologically. The exclusion criteria included 1) patients with non-prostate cancer and benign prostatic hyperplasia; 2) patients without pathological reports; 3) patients with incomplete clinical data; and 4) patients who had received preoperative radiotherapy, chemotherapy or endocrine treatment. Forty-seven samples from patients with transurethral electrocision of the prostate and a pathological diagnosis of prostate cancer were included in the experimental group. In the experimental group, the patients’ ages ranged from 55 to 91, and the average age was 71.15 ± 9.60 years old. In addition, 20 patients with benign prostatic hyperplasia were selected as the control group. In the control group, the patients’ ages ranged from 48 to 79, and the average age was 69.63 ± 8.24 years old. All specimens were from patients who were admitted to the Urology Department of the First Hospital Affiliated of Guangxi Medical University after transurethral electrocision of the prostate. The clinical data collected from all patients included general information  and the results of auxiliary examination, including the preoperative serum PSA level, tumor node metastasis (TNM) staging, pathological findings (including Gleason score), and treatment.Cav-1 protein was detected by the standard streptavidin-peroxidase (SP) immunohistochemical staining method. The tissue slices were processed and incubated with a rabbit anti-human caveolin-1 polyclonal antibody  overnight at 4 °C; then, the slices were incubated with biotin-labeled goat anti-rabbit IgG  at room temperature for 20 min; next, the slices were incubated with peroxidase-labeled streptavidin at room temperature for 20 min and stained with a Tris-HCl solution containing 0.02% 3,9-diaminobenzidine for 5 to 7 min; finally, the slices were stained with hematoxylin, washed with water, dehydrated, removed, sealed, detected and photographed under a microscope.Positive results were analyzed using an Olympus optical microscope, and double-blind randomized observations were made by two chief physicians. Positive caveolin-1 expression was located in the cell membrane and cytoplasm and was indicated by yellowish-brown or brown granules. The scores were calculated according to the staining intensity of positive-staining cells and the number of positive-staining cells (percentage of positive cells) in the tissue slice. (1) The staining intensity was calculated as follows: negative - 0 points; weak positive - 1 point; positive - 2 points; and strong positive - 3 points. For the tissue slices, the staining intensity was calculated as follows: colorless - 0 points; light yellow - 1 point; yellowish-brown - 2 points; and brown - 3 points. (2) The percentage of positive cells were calculated as follows: < 5% - 0 points; ≥ 5 to 25% - 1 point; > 25 to 50% - 2 points; > 50 to 75% - 3 points; and > 75% - 4 points. The final score was calculated by multiplying the staining intensity score by the positive cell percentage. When the final score was ≥4 points, it was considered positive; when the final score was < 4 points, it was considered negative.p < 0.05.All statistical analyses were performed using the SPSS 16.0 software . Group comparisons were performed using the Chi-square test. Differences were considered statistically significant when As shown in Table Caveolin-1 positive staining was found in the cell membrane and cytoplasm of prostate cancer (PCa) cells, presenting as yellowish-brown or brown particles. In 47 cases of prostate cancer, positive expression was found in 29 cases, and negative expression was found in 18 cases; the positive expression rate was 61.70%. In 20 cases of benign prostatic hyperplasia, positive expression was found in 2 cases, and negative expression was found in 18 cases; the positive expression rate was 10.0% .The positive rates of caveolin-1 expression in T1-T2 and T3-T4 PCa tissues were 46.15 and 80.95%, respectively. The expression rate of caveolin-1 increased with the clinical stage. The difference between the two groups was statistically significant (Table P > 0.05)  were 16.67, 68.18 and 68.42%, respectively. The difference was not statistically significant P > 0.0 , the positive rates of expression were 16.67, 68.18 and 68.42%, respectively; there were no significant differences among the three groups. It is assumed that the high expression of caveolin-1 in prostate cancer may be related to the interaction between the phosphorylation and dephosphorylation of scaffolding and multiple signaling pathways and to gene mutation and promoter methylation, but it is not related to serum PSA levels; these results suggest that caveolin-1 may play an important role in the progression of prostate cancer. The results from Mohammed, D.A.  also sugHow does caveolin-1 lose its inhibitory function as a tumor suppressor and instead promote tumor growth? It is generally believed that the mechanism of action may have three points: 1) point mutations ; 2) tyroIn this study, it was found that high caveolin-1 expression is closely related to prostate cancer after transurethral surgery and has a role in promoting tumorigenesis, development, and infiltration.In conclusion, caveolin-1 is highly expressed in prostate cancer and is closely related to the pathological grade and clinical stage of prostate cancer. It may be a novel tumor marker for prostate cancer and has potential clinical value. The expression of caveolin-1 is not related to preoperative serum PSA levels.